The Scientific World Journal: Toxicology The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. Gossypol Promotes Degeneration of Ovarian Follicles in Rats Wed, 23 Jul 2014 10:06:50 +0000 The present study aimed to determine if gossypol interferes with ovarian follicles in rats. Twenty-four female Wistar rats were assigned to two equal groups: one control group and the other dosed with gossypol (25 mg/kg/day, subcutaneously) for 15 days. Ovarian follicles were histologically classified according to the stage of development and as normal or atretic. Gossypol treatment reduced the length of estrous with an increase in the duration of the diestrus phase. This compound was responsible for reduced serum levels of T4 and progesterone. Treatment with gossypol was responsible for a significant reduction in the number of normal ovarian follicles and a significant increase in the number of atretic follicles, both in all stages of development. Thus, treatment of rats with gossypol was responsible for reduction in the number of viable follicles and changes in hormone levels that resulted in interference of the estrous cycle. Ivana Cristina Nunes Gadelha, Michelly Fernandes de Macedo, Sílvia Catarina Salgado Oloris, Marilia Martins Melo, and Benito Soto-Blanco Copyright © 2014 Ivana Cristina Nunes Gadelha et al. All rights reserved. Lymphocyte Oxidative Stress/Genotoxic Effects Are Related to Serum IgG and IgA Levels in Coke Oven Workers Sun, 20 Jul 2014 07:24:59 +0000 We investigated oxidative stress/genotoxic effects levels, immunoglobulin levels, polycyclic aromatic hydrocarbons (PAHs) levels exposed in 126 coke oven workers and in 78 control subjects, and evaluated the association between oxidative stress/genotoxic effects levels and immunoglobulin levels. Significant differences were observed in biomarkers, including 1-hydroxypyrene levels, employment time, percentages of alcohol drinkers, MDA, 8-OHdG levels, CTL levels and CTM, MN, CA frequency, and IgG, IgA levels between the control and exposed groups. Slightly higher 1-OHP levels in smoking users were observed. For the dose-response relationship of IgG, IgA, IgM, and IgE by 1-OHP, each one percentage increase in urinary 1-OHP generates a 0.109%, 0.472%, 0.051%, and 0.067% decrease in control group and generates a 0.312%, 0.538%, 0.062%, and 0.071% decrease in exposed group, respectively. Except for age, alcohol and smoking status, IgM, and IgE, a significant correlation in urinary 1-OHP and other biomarkers in the total population was observed. Additionally, a significant negative correlation in genotoxic/oxidative damage biomarkers of MDA, 8-OH-dG, CTL levels, and immunoglobins of IgG and IgA levels, especially in coke oven workers, was found. These data suggest that oxidative stress/DNA damage induced by PAHs may play a role in toxic responses for PAHs in immunological functions. Meili Gao, Yongfei Li, Aqun Zheng, Xiaochang Xue, Lan Chen, and Yu Kong Copyright © 2014 Meili Gao et al. All rights reserved. Arsenic Modulates Posttranslational S-Nitrosylation and Translational Proteome in Keratinocytes Tue, 08 Jul 2014 00:00:00 +0000 Arsenic is a class I human carcinogen (such as inducing skin cancer) by its prominent chemical interaction with protein thio (-SH) group. Therefore, arsenic may compromise protein S-nitrosylation by competing the -SH binding activity. In the present study, we aimed to understand the influence of arsenic on protein S-nitrosylation and the following proteomic changes. By using primary human skin keratinocyte, we found that arsenic treatment decreased the level of protein S-nitrosylation. This was coincident to the decent expressions of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS). By using LC-MS/MS, around twenty S-nitrosoproteins were detected in the biotin-switched eluent. With the interest that arsenic not only regulates posttranslational S-nitrosylation but also separately affects protein’s translation expression, we performed two-dimensional gel electrophoresis and found that 8 proteins were significantly decreased during arsenic treatment. Whether these decreased proteins are the consequence of protein S-nitrosylation will be further investigated. Taken together, these results provide a finding that arsenic can deplete the binding activity of NO and therefore reduce protein S-nitrosylation. Bin Huang, Kuo-Hao Chiang, Hsin-Su Yu, Ying-Lun Chen, Huey-Ling You, and Wei-Ting Liao Copyright © 2014 Bin Huang et al. All rights reserved. Expression of a Splice Variant of CYP26B1 in Betel Quid-Related Oral Cancer Mon, 07 Jul 2014 09:48:33 +0000 Betel quid (BQ) is a psychostimulant, an addictive substance, and a group 1 carcinogen that exhibits the potential to induce adverse health effects. Approximately, 600 million users chew a variety of BQ. Areca nut (AN) is a necessary ingredient in BQ products. Arecoline is the primary alkaloid in the AN and can be metabolized through the cytochrome P450 (CYP) superfamily by inducing reactive oxygen species (ROS) production. Full-length CYP26B1 is related to the development of oral pharyngeal cancers. We investigated whether a splice variant of CYP26B1 is associated with the occurrence of ROS related oral and pharyngeal cancer. Cytotoxicity assays were used to measure the effects of arecoline on cell viability in a dose-dependent manner. In vitro and in vivo studies were conducted to evaluate the expression of the CYP26B1 splice variant. The CYP26B1 splice variant exhibited lower expression than did full-length CYP26B1 in the human gingival fibroblast-1 and Ca9-22 cell models. Increased expression of the CYP26B1 splice variant was observed in human oral cancer tissue compared with adjacent normal tissue, and increased expression was observed in patients at a late tumor stage. Our results suggested that the CYP26B1 splice variant is associated with the occurrence of BQ-related oral cancer. Ping-Ho Chen, Ka-Wo Lee, Cheng-Chieh Hsu, Jeff Yi-Fu Chen, Yan-Hsiung Wang, Ker-Kong Chen, Hui-Min David Wang, Hurng-Wern Huang, and Bin Huang Copyright © 2014 Ping-Ho Chen et al. All rights reserved. In Vitro Cadmium-Induced Alterations in Growth and Oxidative Metabolism of Upland Cotton (Gossypium hirsutum L.) Wed, 11 Jun 2014 14:13:16 +0000 Cadmium (Cd) is a toxic pollutant, which cause both dose- and time-dependent physiological and biochemical alterations in plants. The present in vitro study was undertaken to explore Cd-induced physiological and biochemical changes in cotton callus culture at 0, 550, 700, 850, and 1000 μM Cd for four different stress periods (7, 14, 21, and 28 days). At 1000 μM Cd, mean growth values were lower than their respective control. The cell protein contents decreased only after 7-day and 14-day stress treatment. At 550 μM Cd, malondialdehyde (MDA) contents decreased after various stress periods except 21-day period. Superoxide dismutase (SOD) activity at 1000 μM Cd improved relative to its respective controls in the first three stress regimes. Almost a decreasing trend in the hydrogen peroxide (H2O2) and peroxidase (POD) activities at all Cd levels after different stress periods was noticed. Ascorbate peroxidase (APX) activity descended over its relevant controls in the first three stress regimes except at 700 μM Cd after 14- and 21-day stress duration. Moreover, catalase (CAT) mean values significantly increased as a whole. From this experiment, it can be concluded that lipid peroxidation as well as reactive oxygen species (ROS) production was relatively higher as has been revealed by higher MDA contents and greater SOD, CAT activities. M. K. Daud, Lei Mei, Ullah Najeeb, Muhammad Azim Khan, Farah Deeba, Irum Raza, Aliya Batool, and S. J. Zhu Copyright © 2014 M. K. Daud et al. All rights reserved. Assessment of Pesticide Residues in Flesh of Catla catla from Ravi River, Pakistan Thu, 05 Jun 2014 08:51:03 +0000 The levels of dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene (DDE), endosulfan, endosulfan sulfate, carbofuran, and cartap which were estimated in the flesh of Catla catla sampled from ten sites of Ravi River between its stretches from Shahdara to Head Balloki were studied to know the level of contamination of the selected pesticides by GC-ECD method. All fish samples were found contaminated with different concentrations of DDT, DDE, endosulfan, and carbofuran; however, DDT and DDE concentrations were more than the maximum residue limits (MRLs) about food standards, while endosulfan sulfate and cartap were not detected. Pesticide concentrations in the fish flesh were ranged from 3.240 to 3.389 for DDT, 2.290 to 2.460 for DDE, 0.112 to 0.136 for endosulfan, and 0.260 to 0.370 μg g−1 for carbofuran. The findings revealed that the pesticide concentrations in the fish flesh decreased in the order: DDT > DDE > carbofuran > endosulfan. After Degh fall and After Hudiara nulla fall river sampling sites were found severely contaminated. It is proposed that a constant monitoring programs are needed to be initiated to overcome the present alarming situation. Mobeen Akhtar, Shahid Mahboob, Salma Sultana, Tayyaba Sultana, Khalid Abdullah Alghanim, and Zubair Ahmed Copyright © 2014 Mobeen Akhtar et al. All rights reserved. Caffeic Acid Phenethyl Ester as a Protective Agent against Nephrotoxicity and/or Oxidative Kidney Damage: A Detailed Systematic Review Tue, 03 Jun 2014 12:04:51 +0000 Caffeic acid phenethyl ester (CAPE), an active component of propolis, has been attracting the attention of different medical and pharmaceutical disciplines in recent years because of its antioxidant, anti-inflammatory, antiproliferative, cytotoxic, antiviral, antifungal, and antineoplastic properties. One of the most studied organs for the effects of CAPE is the kidney, particularly in the capacity of this ester to decrease the nephrotoxicity induced by several drugs and the oxidative injury after ischemia/reperfusion (I/R). In this review, we summarized and critically evaluated the current knowledge regarding the protective effect of CAPE in nephrotoxicity induced by several special medicines such as cisplatin, doxorubicin, cyclosporine, gentamycin, methotrexate, and other causes leading to oxidative renal injury, namely, I/R models and senility. Sumeyya Akyol, Veli Ugurcu, Aynur Altuntas, Rukiye Hasgul, Ozlem Cakmak, and Omer Akyol Copyright © 2014 Sumeyya Akyol et al. All rights reserved. A Chinese Herbal Medicine, Jia-Wei-Xiao-Yao-San, Prevents Dimethylnitrosamine-Induced Hepatic Fibrosis in Rats Tue, 03 Jun 2014 06:43:35 +0000 Jia-wei-xiao-yao-san (JWXYS) is a traditional Chinese herbal medicine that is widely used to treat neuropsychological disorders. Only a few of the hepatoprotective effects of JWXYS have been studied. The aim of this study was to investigate the hepatoprotective effects of JWXYS on dimethylnitrosamine- (DMN-) induced chronic hepatitis and hepatic fibrosis in rats and to clarify the mechanism through which JWXYS exerts these effects. After the rats were treated with DMN for 3 weeks, serum glutamic oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) levels were significantly elevated, whereas the albumin level decreased. Although DMN was continually administered, after the 3 doses of JWXYS were orally administered, the SGOT and SGPT levels significantly decreased and the albumin level was significantly elevated. In addition, JWXYS treatment prevented liver fibrosis induced by DMN. JWXYS exhibited superoxide-dismutase-like activity and dose-dependently inhibited DMN-induced lipid peroxidation and xanthine oxidase activity in the liver of rats. Our findings suggest that JWXYS exerts antifibrotic effects against DMN-induced chronic hepatic injury. The possible mechanism is at least partially attributable to the ability of JWXYS to inhibit reactive-oxygen-species-induced membrane lipid peroxidation. Shu-Chen Chien, Wei-Chiao Chang, Pu-Hua Lin, Wei-Pin Chang, Shih-Chung Hsu, Jung-Chen Chang, Ya-Chieh Wu, Jin-Kuo Pei, and Chia-Hsien Lin Copyright © 2014 Shu-Chen Chien et al. All rights reserved. Hydrogen Sulfide Increases Nitric Oxide Production and Subsequent S-Nitrosylation in Endothelial Cells Wed, 21 May 2014 06:17:17 +0000 Hydrogen sulfide (H2S) and nitric oxide (NO), two endogenous gaseous molecules in endothelial cells, got increased attention with respect to their protective roles in the cardiovascular system. However, the details of the signaling pathways between H2S and NO in endothelia cells remain unclear. In this study, a treatment with NaHS profoundly increased the expression and the activity of endothelial nitric oxide synthase. Elevated gaseous NO levels were observed by a novel and specific fluorescent probe, 5-amino-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid methyl ester (FA-OMe), and quantified by flow cytometry. Further study indicated an increase of upstream regulator for eNOS activation, AMP-activated protein kinase (AMPK), and protein kinase B (Akt). By using a biotin switch, the level of NO-mediated protein S-nitrosylation was also enhanced. However, with the addition of the NO donor, NOC-18, the expressions of cystathionine-γ-lyase, cystathionine-β-synthase, and 3-mercaptopyruvate sulfurtransferase were not changed. The level of H2S was also monitored by a new designed fluorescent probe, 4-nitro-7-thiocyanatobenz-2-oxa-1,3-diazole (NBD-SCN) with high specificity. Therefore, NO did not reciprocally increase the expression of H2S-generating enzymes and the H2S level. The present study provides an integrated insight of cellular responses to H2S and NO from protein expression to gaseous molecule generation, which indicates the upstream role of H2S in modulating NO production and protein S-nitrosylation. Ping-Ho Chen, Yaw-Syan Fu, Yun-Ming Wang, Kun-Han Yang, Danny Ling Wang, and Bin Huang Copyright © 2014 Ping-Ho Chen et al. All rights reserved. Progesterone Increases Apoptosis and Inversely Decreases Autophagy in Human Hepatoma HA22T/VGH Cells Treated with Epirubicin Mon, 19 May 2014 07:19:09 +0000 Hepatocellular carcinoma (HCC) is the leading cause of cancer-related deaths worldwide. Epirubicin can induce intracellular reactive oxygen species and is widely used to treat unresectable HCC. Progesterone has been found to inhibit the proliferation of hepatoma cells. This study was designed to test the combined effects of epirubicin and progesterone on human hepatoma cell line, HA22T/VGH. These cells were treated with different concentrations of epirubicin with or without the coaddition of 30 μM progesterone and then analyzed for apoptosis, autophagy, and expressions of apoptotic-related proteins and multidrug-resistant gene. Epirubicin treatment dose-dependently inhibited the growth of HA22T/VGH cells. Addition of 30 μM progesterone, which was inactive alone, augmented the effect of epirubicin on the inhibition of growth of HA22T/VGH cells. Cotreatment with progesterone enhanced epirubicin-induced apoptosis, as evidenced by greater increase in caspase-3 activity and in the ratio of the apoptosis-regulating protein, Bax/Bcl-XL. The combination also caused a decrease in autophagy and in the expression of multidrug resistance-related protein 1 mRNA compared to epirubicin alone. This study shows the epirubicin/progesterone combination was more effective in increasing apoptosis and inversely decreasing autophagy on HA22T/VGH cells treated with epirubicin alone, suggesting that this combination can potentially be used to treat HCC. Wen-Tsan Chang, Hsiao-Ling Cheng, Bau-Shan Hsieh, Chien-Chih Chiu, King-Teh Lee, and Kee-Lung Chang Copyright © 2014 Wen-Tsan Chang et al. All rights reserved. Interval between Intra-Arterial Infusion Chemotherapy and Surgery for Locally Advanced Oral Squamous Cell Carcinoma: Impacts on Effectiveness of Chemotherapy and on Overall Survival Sun, 18 May 2014 11:47:34 +0000 Background. The interval between intra-arterial infusion chemotherapy (IAIC) and surgery was investigated in terms of its effects on survival in patients with locally advanced oral squamous cell carcinoma (OSCC). Methods. This retrospective study analyzed 126 patients who had completed treatment modalities for stage IV OSCC. All patients were followed up for 3 years. Kaplan-Meier and Cox regression methods were used to determine how survival was affected by general factors, primary tumor volume, TNM stage, and duration of neoadjuvant chemotherapy. Results. In 126 patients treated for locally advanced OSCC by preoperative induction IAIC using methotrexate, multivariate analysis of relevant prognostic factors showed that an IAIC duration longer than 90 days was significantly associated with poor prognosis (hazard ratio, 1.77; ). Conclusions. Duration of IAIC is a critical factor in the effectiveness of multimodal treatment for locally advanced OSCC. Limiting the induction course to 90 days improves overall survival. Chih-Fung Wu, Chien-Hsing Lee, Edward Hsi, Chung-Ho Chen, and Jen-Yang Tang Copyright © 2014 Chih-Fung Wu et al. All rights reserved. Caenorhabditis elegans as Model System in Pharmacology and Toxicology: Effects of Flavonoids on Redox-Sensitive Signalling Pathways and Ageing Wed, 30 Apr 2014 10:48:47 +0000 Flavonoids are secondary plant compounds that mediate diverse biological activities, for example, by scavenging free radicals and modulating intracellular signalling pathways. It has been shown in various studies that distinct flavonoid compounds enhance stress resistance and even prolong the life span of organisms. In the last years the model organism C. elegans has gained increasing importance in pharmacological and toxicological sciences due to the availability of various genetically modified nematode strains, the simplicity of modulating genes by RNAi, and the relatively short life span. Several studies have been performed demonstrating that secondary plant compounds influence ageing, stress resistance, and distinct signalling pathways in the nematode. Here we present an overview of the modulating effects of different flavonoids on oxidative stress, redox-sensitive signalling pathways, and life span in C. elegans introducing the usability of this model system for pharmacological and toxicological research. Karoline Koch, Susannah Havermann, Christian Büchter, and Wim Wätjen Copyright © 2014 Karoline Koch et al. All rights reserved. Mineral Element Contents in Commercially Valuable Fish Species in Spain Wed, 23 Apr 2014 13:52:04 +0000 The aim of this study was to measure selected metal concentrations in Trachurus trachurus, Trachurus picturatus, and Trachurus mediterraneus, which are widely consumed in Spain. Principal component analysis suggested that the variable Cr was the main responsible variable for the identification of T. trachurus, the variables As and Sn for T. mediterraneus, and the rest of variables for T. picturatus. This well-defined discrimination between fish species provided by mineral element allows us to distinguish them on the basis of their metal content. Based on the samples collected, and recognizing the inferential limitation of the sample size of this study, the metal concentrations found are below the proposed limit values for human consumption. However, it should be taken into consideration that there are other dietary sources of these metals. In conclusion, metal contents in the fish species analyzed are acceptable for human consumption from a nutritional and toxicity point of view. Ana Rivas, Luis Peña-Rivas, Eduardo Ortega, Concepción López-Martínez, Fátima Olea-Serrano, and Maria Luisa Lorenzo Copyright © 2014 Ana Rivas et al. All rights reserved. Evaluation on Genotoxicity and Teratogenicity of Aqueous Extract from Cyclocarya paliurus Leaves Tue, 18 Mar 2014 07:14:39 +0000 Tremendous attentions have been attracted to the foods labeled with natural, green, organic, and nuisanceless conception of healthy diet. Therefore, it is of great significance to establish relative defining guidance for safe assessment of botanicals. Cyclocarya paliurus (Batal.) Iljinsk (family Cyclocaryaceae), called sweet tea tree, is a well-known edible and medicinal plant, which has been widely used in China as drug formulation for the treatment of hypertension and diabetes. Despite its benefits, no reports have been described on the safe assessment of C. paliurus leaves aqueous extract. In this study, we have conducted the genotoxicity assay (including Ames test, bone marrow polychromatic erythrocyte micronucleus test, and sperm abnormality test in mice) and traditional teratogenicity assay in rats (maternal toxicity, embryo toxicity, and teratogenicity test) to assess the genetic and teratogenic safety of aqueous extracts from C. paliurus leaves. Results of each assay show that the highest dose of C. paliurus leaves aqueous extract is considered relatively nonmutagenic and nonteratogenic, revealing that C. paliurus leaves possess safety and quality as a functional additional ingredient in food. Lihong Deng, Jiandu Lei, Jing He, Jing Liu, Luying Wang, Rui Zhang, Xinhua Liu, and Yun Liu Copyright © 2014 Lihong Deng et al. All rights reserved. Caloric Restriction Prevents Lead-Induced Oxidative Stress and Inflammation in Rat Liver Thu, 20 Feb 2014 09:43:58 +0000 The aim of the present study is to investigate the effects of caloric restriction on liver of lead-administered rat. Male Sprague-Dawley rats were randomly divided into two groups: Ad libitum fed group (AL, free access to normal rat chow) and caloric restriction group (CR, fed 65% of AL animals’ food intake). After 6 weeks, half of the animals of each group were injected lead acetate and the other half were injected saline. Liver tissue samples were collected at the end of the experiments. Glutathione peroxidase (GPx), superoxide dismutase (SOD), malondialdehyde (MDA), and tumor necrosis factor (TNF-α) were measured in the tissue extracts. Histological studies were also performed. Our results showed that lead administrations (not saline injections) reduced liver SOD and GPx and increased MDA and TNF-α in AL animals, but in the CR animals lead injections did not significantly change the measured parameters. The histological studies supported the biochemical findings. We concluded that 65% CR may prevent lead-induced oxidative stress and inflammation in rat liver. Mustafa Mohammadi, Rana Ghaznavi, Rana Keyhanmanesh, Hamid Reza Sadeghipour, Roya Naderi, and Hossein Mohammadi Copyright © 2014 Mustafa Mohammadi et al. All rights reserved. Effect of Fluorosis on Liver Cells of VC Deficient and Wild Type Mice Sun, 16 Feb 2014 07:56:04 +0000 For decades, mouse and other rodents have been used for the study of oxidative or related studies such as the effect of fluoride. It is known that rodents normally synthesize their own vitamin C (VC) due to the presence of a key enzyme in ascorbic acid synthesis, l-gulono-lactone-γ-oxidase (Gulo), while humans do not have the capacity of VC synthesis due to the deletion of most parts of the GULO gene. The spontaneous fracture (sfx) mouse recently emerged as a model for study of VC deficiency. We investigated the effect of fluoride on liver cells from wild type Balb/c and sfx mice. We found that activities of SOD, GPx, and CAT were reduced in both wild type and sfx mice; however, the amount of reduction in the sfx cells is more than that in Balb/c cells. In addition, while both cells increased MDA, the increase in the sfx cells is greater than that in Balb/c cells. Gene networks of Sod, Gpx, and Cat in the liver of humans and mice are also different. Our study suggests that reaction to fluoride in vitamin C deficient mice might be different from that of wild type mice. Wei Wei, Yan Jiao, Yonghui Ma, John M. Stuart, Xiudian Li, Fusheng Zhao, Lishi Wang, DianJun Sun, and Weikuan Gu Copyright © 2014 Wei Wei et al. All rights reserved. Assessment of Cellular Responses after Short- and Long-Term Exposure to Silver Nanoparticles in Human Neuroblastoma (SH-SY5Y) and Astrocytoma (D384) Cells Thu, 13 Feb 2014 12:01:15 +0000 Silver nanoparticle (AgNP, 20 nm) neurotoxicity was evaluated by an integrated in vitro testing protocol employing human cerebral (SH-SY5Y and D384) cell lines. Cellular response after short-term (4–48 h, 1–100 μg/ml) and prolonged exposure (up to 10 days, 0.5–50 μg/ml) to AgNP was assessed by MTT, calcein-AM/PI, clonogenic tests. Pulmonary A549 cells were employed for data comparison along with silver nitrate as metal ionic form. Short-term data: (i) AgNP produced dose- and time-dependent mitochondrial metabolism changes and cell membrane damage (effects starting at 25 μg/ml after 4 h: EC50s were 40.7 ± 2.0 and 49.5 ± 2.1 μg/ml for SH-SY5Y and D384, respectively). A549 were less vulnerable; (ii) AgNP doses of ≤ 18 μg/ml were noncytotoxic; (iii) AgNO3 induced more pronounced effects compared to AgNP on cerebral cells. Long-term data: (i) low AgNP doses (≤1 μg/ml) compromised proliferative capacity of all cell types (cell sensibility: SHSY5Y > A549 > D384). Colony number decrease in SH-SY5Y and D384 was 50% and 25%, respectively, at 1 μg/ml, and lower dose (0.5 μg/ml) was significantly effective towards SH-SY5Y and pulmonary cells; (ii) cell proliferation activity was more affected by AgNO3 than AgNPs. In summary, AgNP-induced cytotoxic effects after short-term and prolonged exposure (even at low doses) were evidenced regardless of cell model types. Teresa Coccini, Luigi Manzo, Vittorio Bellotti, and Uliana De Simone Copyright © 2014 Teresa Coccini et al. All rights reserved. Potent Antioxidative and UVB Protective Effect of Water Extract of Eclipta prostrata L. Sun, 09 Feb 2014 13:45:34 +0000 Oxidative stress, including Ultraviolet (UV) irradiation-induced skin damage, is involved in numerous diseases. This study demonstrates that water extract of Eclipta prostrata L. (WEP) has a potent effect in scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide radicals, and chelating ferrous ion, exhibiting IC50 values of 0.23 mg/mL, 0.48 mg/mL, and 1.25 mg/mL, respectively. The WEP total phenol content was 176.45 mg gallic acid equivalents (GAE)/g sample. Chlorogenic acid, a component of the plant's active ingredients, was determined by HPLC and antioxidative assay. However, no caffeic acid, stigmasterol, or wedelolactone was present in WEP. WEP absorbs both UVA and UVB irradiation, and furthermore, the extract shows a dose-dependent response in the protection of HaCaT human keratinocytes and mouse fibroblasts 3T3 cells against UVB-induced cytotoxicity, which may result from a synergistic effect between chlorogenic acid and other active components present in WEP. Chin-Feng Chan, Wen-Ying Huang, Hong-Yi Guo, and Bo Rong Wang Copyright © 2014 Chin-Feng Chan et al. All rights reserved. Diverse Effects of Lead Nitrate on the Proliferation, Differentiation, and Gene Expression of Stem Cells Isolated from a Dental Origin Tue, 28 Jan 2014 08:27:29 +0000 Lead (Pb2+) exposure continues to be a significant public health problem. Therefore, it is vital to have a continuous epidemiological dataset for a better understanding of Pb2+ toxicity. In the present study, we have exposed stem cells isolated from deciduous and permanent teeth, periodontal ligament, and bone marrow to five different types of Pb2+ concentrations (160, 80, 40, 20, and 10 µM) for 24 hours to identify the adverse effects of Pb2+ on the proliferation, differentiation, and gene expression on these cell lines. We found that Pb2+ treatment altered the morphology and adhesion of the cells in a dose-dependent manner. There were no significant changes in terms of cell surface phenotypes. Cells exposed to Pb2+ continued to differentiate into chondrogenesis and adipogenesis, and a severe downregulation was observed in osteogenesis. Gene expression studies revealed a constant expression of key markers associated with stemness (Oct 4, Rex 1) and DNA repair enzyme markers, but downregulation occurred with some ectoderm and endoderm markers, demonstrating an irregular and untimely differentiation trail. Our study revealed for the first time that Pb2+ exposure not only affects the phenotypic characteristics but also induces significant alteration in the differentiation and gene expression in the cells. Mariam Abdullah, Fazliny Abd. Rahman, Nareshwaran Gnanasegaran, Vijayendran Govindasamy, Noor Hayaty Abu Kasim, and Sabri Musa Copyright © 2014 Mariam Abdullah et al. All rights reserved. Toxicity Effect of Silver Nanoparticles in Brine Shrimp Artemia Thu, 02 Jan 2014 08:41:25 +0000 The present study revealed the toxic effect of silver nanoparticles (AgNPs) in Artemia nauplii and evaluated the mortality rate, hatching percentage, and genotoxic effect in Artemia nauplii/cysts. The AgNPs were commercially purchased and characterized using field emission scanning electron microscope with energy dispersive X-ray spectroscopy. Nanoparticles were spherical in nature and with size range of 30–40 nm. Artemia cysts were collected from salt pan, processed, and hatched in sea water. Artemia nauplii (II instar) were treated using silver nanoparticles of various nanomolar concentrations and LC50 value (10 nM) and mortality rate (24 and 48 hours) was evaluated. Hatching percentage of decapsulated cysts treated with AgNPs was examined. Aggregation of AgNPs in the gut region of nauplii was studied using phase contrast microscope and apoptotic cells in nauplii stained with acridine orange were observed using fluorescence microscope. DNA damage of single cell of nauplii was determined by comet assay. This study showed that as the concentration of AgNPs increased, the mortality rate, aggregation in gut region, apoptotic cells, and DNA damage increased in nauplii, whereas the percentage of hatching in Artemia cysts decreased. Thus this study revealed that the nanomolar concentrations of AgNPs have toxic effect on both Artemia nauplii and cysts. Chinnasamy Arulvasu, Samou Michael Jennifer, Durai Prabhu, and Devakumar Chandhirasekar Copyright © 2014 Chinnasamy Arulvasu et al. All rights reserved. Accumulation of Scandium in Plasma in Patients with Chronic Renal Failure Thu, 12 Dec 2013 16:56:56 +0000 Scandium (Sc) is an element with many industrial applications, but relatively little is known about its physiological and/or toxicological effects, and very little data are available concerning the role of Sc in chronic renal failure (CRF). This paper examines the changes in plasma levels of Sc in predialysis patients with CRF and the relationship with blood parameters. The participants in this trial were 48 patients with CRF in predialysis and 53 healthy controls. Erythrocyte, haemoglobin, and haematocrit counts in blood were determined, and levels of creatinine, urea, uric acid, albumin, total protein and Sc were measured in plasma. The glomerular filtration rate (GFR) was calculated using the Cockcroft-Gault index. The CRF patients were found to have higher plasma levels of creatinine, urea, uric acid, albumin, total protein, and Sc and a lower GFR than that the controls. Scandium in plasma was positively correlated with creatinine and plasma urea and negatively correlated with GFR, haemoglobin, and haematocrit and was associated with the risk of lower levels of erythrocytes, haemoglobin, and haematocrit. CRF was associated with increases in the circulating levels of scandium. Cristina Sánchez-González, Carlos López-Chaves, Lorenzo Rivas-García, Pilar Galindo, Jorge Gómez-Aracena, Pilar Aranda, and Juan Llopis Copyright © 2013 Cristina Sánchez-González et al. All rights reserved. Metal Concentrations in Cosmetics Commonly Used in Nigeria Thu, 05 Dec 2013 15:46:49 +0000 Trace amounts of potentially toxic metals can be either intentionally added to cosmetics or present as impurities in the raw materials. In the present study, the levels of lead, cadmium, nickel, chromium, and mercury have been assessed in 28 body creams and lotions, 10 powders, 3 soaps, 5 eye make-ups, and 4 lipsticks widely available on Nigerian markets. The increases over suggested or mandated levels of lead in these creams and lotions ranged from 6.1 to 45.9 and from 1.2 to 9.2 mg kg−1 when compared with Cosmetic Ingredients Review Expert Panel 2007 and German safe maximum permissible limit of lead in cosmetics, respectively. About 61% of the body cosmetics, the lotions, and the creams contained detectable levels of nickel ranging from 1.1 to 6.4–9.2 mg kg−1. Chromium and mercury were undetected in 100% of the cosmetic product. Taken together, lead and cadmium were high in creams and lotions. Most of the imported creams and creamy white coloured cosmetics contained higher levels of metal contaminants than the other colours. Regulatory Agencies in developing nations should take appropriate action for cosmetics that contain lead and cadmium beyond the reference limits. Orish Ebere Orisakwe and Jonathan Oye Otaraku Copyright © 2013 Orish Ebere Orisakwe and Jonathan Oye Otaraku. All rights reserved. Drinking Water Fluoride Levels for a City in Northern Mexico (Durango) Determined Using a Direct Electrochemical Method and Their Potential Effects on Oral Health Wed, 20 Nov 2013 14:04:09 +0000 Fluoride is ingested primarily through consuming drinking water. When drinking water contains fluoride concentrations >0.7 parts per million (ppm), consuming such water can be toxic to the human body; this toxicity is called “fluorosis.” Therefore, it is critical to determine the fluoride concentrations in drinking water. The objective of this study was to determine the fluoride concentration in the drinking water of the city of Durango. The wells that supply the drinking water distribution system for the city of Durango were studied. One hundred eighty-nine (189) water samples were analyzed, and the fluoride concentration in each sample was quantified as established by the law NMX-AA-077-SCFI-2001. The fluoride concentrations in such samples varied between 2.22 and 7.23 ppm with a 4.313 ± 1.318 ppm mean concentration. The highest values were observed in the northern area of the city, with a 5.001 ± 2.669 ppm mean value. The samples produced values that exceeded the national standard for fluoride in drinking water. Chronic exposure to fluoride at such concentrations produces harmful health effects, the first sign of which is dental fluorosis. Therefore, it is essential that the government authorities implement water defluoridation programs and take preventative measures to reduce the ingestion of this toxic halogen. Nelly Molina Frechero, Leonor Sánchez Pérez, Enrique Castañeda Castaneira, Anastasio Oropeza Oropeza, Enrique Gaona, José Salas Pacheco, and Ronell Bologna Molina Copyright © 2013 Nelly Molina Frechero et al. All rights reserved. Cooccurrence of Mycotoxins in Maize and Poultry Feeds from Brazil by Liquid Chromatography/Tandem Mass Spectrometry Thu, 14 Nov 2013 08:41:58 +0000 The objective of this study was to quantitatively evaluate mycotoxins in samples of maize and poultry feed produced in Brazil. A multimycotoxin method based on HPLC-MS/MS was applied to investigate the occurrence of toxical fungal metabolites in 119 samples collected from poultry feed factory integrated poultry farms: maize grain (74), poultry feed (36), and feed factory residue (9). Twenty of 101 fungal metabolites investigated were detected and quantified in the samples: aflatoxins B1, B2, G1, and G2, fumonisins B1, B2, and B3, hydrolyzed fumonisin B1, zearalenone, agroclavine, chanoclavine, deoxynivalenol, and nivalenol, and enniatin A, A1, B, B1, beauvericin, kojic acid, and moniliformin. Most samples were contaminated with more than one mycotoxin. All samples were contaminated with fumonisins, with medians values of 1,840 μg/kg, 239 μg/kg, and 23,676 μg/kg for maize, feed, and factory residue samples, respectively. Surprisingly, beauvericin was detected in more than 90% of samples. The median contaminations of aflatoxin and trichothecenes were low, near LOD values. The factory residue presented highest contamination levels for all mycotoxins. This is the first study dealing with agroclavine, chanoclavine, enniatin A, A1, B, B1, beauvericin, and kojic acid contamination of maize and poultry feeds from Brazil. Maria de Lourdes Mendes de Souza, Michael Sulyok, Otniel Freitas-Silva, Sônia Soares Costa, Catherine Brabet, Miguel Machinski Junior, Beatriz Leiko Sekiyama, Eugenia Azevedo Vargas, Rudolf Krska, and Rainer Schuhmacher Copyright © 2013 Maria de Lourdes Mendes de Souza et al. All rights reserved. Relevance of Biofilms in the Pathogenesis of Shiga-Toxin-Producing Escherichia coli Infection Tue, 12 Nov 2013 14:56:24 +0000 The present study was designed to determine the relationships among biofilm formation, cellular stress and release of Shiga toxin (Stx) by three different clinical Shiga toxin-producing Escherichia coli (STEC) strains. The biofilm formation was determined using crystal violet stain in tryptic soy broth or thioglycollate medium with the addition of sugars (glucose or mannose) or hydrogen peroxide. The reactive oxygen species (ROSs) were detected by the reduction of nitro blue tetrazolium and reactive nitrogen intermediates (RNI) determined by the Griess assay. In addition, the activities of two antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), were studied. For the cytotoxicity studies, Vero cells were cultured with Stx released of STEC biofilms. The addition of sugars in both culture mediums resulted in an increase in biofilm biomass, with a decrease in ROS and RNI production, low levels of SOD and CAT activity, and minimal cytotoxic effects. However, under stressful conditions, an important increase in the antioxidant enzyme activity and high level of Stx production were observed. The disturbance in the prooxidant-antioxidant balance and its effect on the production and release of Stx evaluated under different conditions of biofilm formation may contribute to a better understanding of the relevance of biofilms in the pathogenesis of STEC infection. Natalia Angel Villegas, José Baronetti, Inés Albesa, Rosana Polifroni, Alberto Parma, Analía Etcheverría, Maria Becerra, Nora Padola, and Maria Paraje Copyright © 2013 Natalia Angel Villegas et al. All rights reserved. Evaluation of Genotoxic Effects of New Molecules with Possible Trypanocidal Activity for Chagas Disease Treatment Thu, 07 Nov 2013 13:42:50 +0000 Chagas disease is responsible for a large number of human infections and many are also at risk of infection. There is no effective drug for Chagas disease treatment. The Institute of Pharmaceutical Technology at Fiocruz, Brazil, has designed three nitro analogs of the nitroimidazole-thiadiazole, megazol: two triazole analogs PTAL 05-02 and PAMT 09 and a pyrazole analog PTAL 04-09. A set of Salmonella enterica serovar Typhimurium strains were used in the bacterial reverse mutation test (Ames test) to determine the mutagenicity and cytotoxicity of megazol and its nitro analogs. Megazol presented positive mutagenic activity at very low concentration, either with or without metabolic activation S9 mix. The mutagenic response of the analogs was detected at higher concentration than the lowest megazol concentration to yield mutagenic activity showing that new advances can be made to develop new analogs. The micronucleus test with rat macrophage cells was used in the genotoxic evaluation. The analogs were capable of inducing micronucleus formation and showed cytotoxic effects. PTAL 04-09 structural modifications might be better suitable for the design of promising new drugs candidate for Chagas’ disease treatment. Francisco V. C. Mello, Alcione S. Carvalho, Mônica M. Bastos, Nubia Boechat, Claudia A. F. Aiub, and Israel Felzenszwalb Copyright © 2013 Francisco V. C. Mello et al. All rights reserved. The Mitigating Effect of Citrullus colocynthis (L.) Fruit Extract against Genotoxicity Induced by Cyclophosphamide in Mice Bone Marrow Cells Thu, 07 Nov 2013 13:09:41 +0000 Possible genoprotective effect of Citrullus colocynthis (L.) (CCT) fruits extract against cyclophosphamide- (CP-)induced DNA damage in mice bone marrow cells was evaluated using micronucleus assay, as an index of induced chromosomal damage. Mice were preadministered with different doses of CCT via intraperitoneal injection for 7 consecutive days followed by injection with CP (70 mg/kg b.w.) 1 hr after the last injection of CCT. After 24 hr, mice were scarified to evaluate the frequency of micronucleated polychromatic erythrocytes (MnPCEs). In addition, the number of polychromatic erythrocytes (PCEs) among 1000 normochromatic erythrocytes (NCEs) per animal was recorded to evaluate bone marrow. Pretreatment with CCT significantly reduced the number of MnPCEs induced by CP in bone marrow cells (). At 200 mg/kg, CCT had a maximum chemoprotective effect and reduced the number of MnPCEs by 6.37-fold and completely normalized the mitotic activity. CCT also led to marked proliferation and hypercellularity of immature myeloid elements after mice were treated with CP and mitigated the bone marrow suppression. Our study revealed that CCT has an antigenotoxic effect against CP-induced oxidative DNA damage in mice. Therefore, it could be used concomitantly as a supplement to protect people undergoing chemotherapy. Mohammad Shokrzadeh, Aroona Chabra, Farshad Naghshvar, and Amirhossein Ahmadi Copyright © 2013 Mohammad Shokrzadeh et al. All rights reserved. Acute Toxicity and Genotoxic Activity of Avocado Seed Extract (Persea americana Mill., c.v. Hass) Tue, 05 Nov 2013 14:50:56 +0000 The use of vegetal extracts requires toxicological and genotoxic evaluations to establish and verify safety before being added to human cosmetic, pharmaceutical medicine, or alimentary products. Persea americana seeds have been used in traditional medicine as treatment for several diseases. In this work, the ethanolic seed extract of Persea americana was evaluated with respect to its genotoxic potential through micronucleus assay in rodents. The frequency of micronuclei in groups of animals treated with avocado seed extract showed no differences compared to the negative control (vehicle); therefore, it is considered that the avocado seed extract showed no genotoxic activity in the micronucleus test. Eduardo Padilla-Camberos, Moisés Martínez-Velázquez, José Miguel Flores-Fernández, and Socorro Villanueva-Rodríguez Copyright © 2013 Eduardo Padilla-Camberos et al. All rights reserved. Genotoxic Potential and Physicochemical Parameters of Sinos River, Southern Brazil Tue, 29 Oct 2013 14:43:11 +0000 The present study aimed to evaluate the physicochemical parameters and the genotoxic potential of water samples collected in the upper, middle, and lower courses of the Sinos River, southern Brazil. The comet assay was performed in the peripheral blood of fish Hyphessobrycon luetkenii exposed under laboratory conditions to water samples collected in summer and winter in three sampling sites of Sinos River. Water quality analysis demonstrated values above those described in Brazilian legislation in Parobé and Sapucaia do Sul sites, located in the middle and in the lower courses of the Sinos River, respectively. The Caraá site, located in the upper river reach, presented all the physicochemical parameters in accordance with the allowed limits in both sampling periods. Comet assay in fish revealed genotoxicity in water samples collected in the middle course site in summer and in the three sites in winter when compared to control group. Thus, the physicochemical parameters indicated that the water quality of the upper course complies with the limits set by the national guidelines, and the ecotoxicological assessment, however, indicated the presence of genotoxic agents. The present study highlights the importance of combining water physicochemical analysis and bioassays to river monitoring. Madalena C. S. Scalon, Ciliana Rechenmacher, Anna Maria Siebel, Michele L. Kayser, Manoela T. Rodrigues, Sharbel W. Maluf, Marco Antonio S. Rodrigues, and Luciano Basso da Silva Copyright © 2013 Madalena C. S. Scalon et al. All rights reserved. Cytotoxic and Apoptotic Effects of Different Extracts of Artemisia turanica Krasch. on K562 and HL-60 Cell Lines Mon, 28 Oct 2013 14:11:28 +0000 Artemisia is an important genus of Iranian flora. Cytotoxic activities for some species of the genus have already been reported. In this study, we have investigated the cytotoxic effects of n-hexane, CH2Cl2, EtOAc, EtOH, and EtOH/H2O (1 : 1) extracts of A. turanica Krasch. on two human leukemic cancer cell lines (K562 and HL-60) and J774 as normal cells using alamarBlue (resazurin) assay. PI staining of the fragmented DNA and western blot analysis were used to evaluate the possible apoptotic effect of the extract. The CH2Cl2 extract of A. turanica showed the most antiproliferative effect on cancer cells among all tested extracts with IC50 values of 69 and 104 μg/mL on K562 and HL-60 cells, respectively, whereas the normal cells were not affected significantly by this extract. Sub-G1 peak in the flow cytometry histogram of the cells treated with CH2Cl2 extract of A. turanica and cleavage of PARP protein confirmed the induction of apoptosis with CH2Cl2 extract. Taken together, the findings of the present work suggest the anticancer potential of CH2Cl2 extract of A. turanica on human leukemic cancer cell lines. Zahra Tayarani-Najaran, Mahla Sareban, Atefeh Gholami, Seyed Ahmad Emami, and Mahdi Mojarrab Copyright © 2013 Zahra Tayarani-Najaran et al. All rights reserved. Genotoxicity Assessment of Multispecies Probiotics Using Reverse Mutation, Mammalian Chromosomal Aberration, and Rodent Micronucleus Tests Wed, 23 Oct 2013 11:22:26 +0000 Genotoxicity assessment is carried out on freeze dried powder of cultured probiotics containing Lactobacillus rhamnosus LCR177, Bifidobacterium adolescentis BA286, and Pediococcus acidilactici PA318. Ames tests, in vitro mammalian chromosome aberration assay, and micronucleus tests in mouse peripheral blood are performed. For 5 strains of Salmonella Typhimurium, the Ames tests show no increased reverse mutation upon exposure to the test substance. In CHO cells, the frequency of chromosome aberration does not increase in responding to the treatment of probiotics. Likewise, the frequency of micronucleated reticulocytes in probiotics-fed mice is indistinguishable from that in the negative control group. Taken together, the toxicity assessment studies suggest that the multispecies probiotic mixture does not have mutagenic effects on various organisms. Yi-Jen Chiu, Mun-Kit Nam, Yueh-Ting Tsai, Chun-Chi Huang, and Cheng-Chih Tsai Copyright © 2013 Yi-Jen Chiu et al. All rights reserved. Voluntary Exercise Prevents Lead-Induced Elevation of Oxidative Stress and Inflammation Markers in Male Rat Blood Thu, 03 Oct 2013 18:45:58 +0000 Regular mild exercise enhances antioxidant and anti-inflammatory systems of the body. The present study investigates voluntary exercise effects on lead toxicity as a known oxidative stressor. Male Sprague-Dawley rats were randomly divided into 2 groups. Sedentary control: the animals were housed 7 weeks in the regular cages. Exercise group: the animals were housed 7 weeks in the running wheel equipped cages, that is, the animal model of voluntary exercise. During the 7th week, all animals were administered lead acetate. Blood samples were collected at the end of the 6th week and 7th week (before and after lead administrations). Glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and tumor necrosis factor (TNF-) were measured in the samples. Our results showed that lead administration reduced blood SOD, GPx and CAT and increased TNF-; in the controls, but in the exercise group, changes were not statistically significant. MDA in both groups increased after lead injections but it was significantly lower in exercise group compared to the sedentary animals. We concluded that voluntary exercise may be considered as a preventive tool against lead-induced oxidative stress and inflammation. Mustafa Mohammadi, Rana Ghaznavi, Rana Keyhanmanesh, Hamid Reza Sadeghipour, Roya Naderi, and Hossein Mohammadi Copyright © 2013 Mustafa Mohammadi et al. All rights reserved. In Vitro Effect of Sodium Fluoride on Malondialdehyde Concentration and on Superoxide Dismutase, Catalase, and Glutathione Peroxidase in Human Erythrocytes Tue, 24 Sep 2013 16:24:22 +0000 The aim of this paper was to describe the in vitro effect of sodium fluoride (NaF) on the specific activity of the major erythrocyte antioxidant enzymes, as well as on the membrane malondialdehyde concentration, as indicators of oxidative stress. For this purpose, human erythrocytes were incubated with NaF (0, 7, 28, 56, and 100 μg/mL) or NaF (100 μg/mL) + vitamin E (1, 2.5, 5 and 10 μg/mL). The malondialdehyde (MDA) concentration on the surface of the erythrocytes was determined, as were the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GlPx). Our results demonstrated that erythrocytes incubated with increasing NaF concentrations had an increased MDA concentration, along with decreased activity of antioxidant enzymes. The presence of vitamin E partially reversed the toxic effects of NaF on erythrocytes. These findings suggest that NaF induces oxidative stress in erythrocytes in vitro, and this stress is partially reversed by the presence of vitamin E. José Gutiérrez-Salinas, Liliana García-Ortíz, José A. Morales González, Sergio Hernández-Rodríguez, Sotero Ramírez-García, Norma R. Núñez-Ramos, and Eduardo Madrigal-Santillán Copyright © 2013 José Gutiérrez-Salinas et al. All rights reserved. Effects of Resveratrol on Methotrexate-Induced Testicular Damage in Rats Sun, 28 Jul 2013 12:03:33 +0000 This study investigated the probable protective effects of resveratrol (RES), an antioxidant, against methotrexate- (MTX-) induced testis damage. Twenty-four male Sprague Dawley rats were randomly divided into four groups: control, RES, MTX, and MTX + RES groups. Rats were sacrificed at the end of the experiment. Plasma and tissue malondialdehyde (MDA) levels, superoxide dismutase (SOD) and catalase (CAT) activity in tissue, testicular histopathological damage scores, and testicular and epididymal epithelial apoptotic index (AI) were evaluated. The MTX group had significantly higher plasma and tissue MDA levels and significantly lower SOD and CAT activity than those of the control group. In the MTX + RES group, plasma and tissue MDA levels decreased significantly and SOD activity rose significantly compared to the MTX group. The MTX group had significantly lower Johnsen’s testicular biopsy score (JTBS) values than those of the control group. JTBS was significantly higher in the MTX + RES group than in the MTX group. AI increased in the testis and epididymis in the MTX group and significantly decreased in the MTX + RES group. Our results indicate that RES has protective effects against MTX-induced testis damage at the biochemical, histopathological, and apoptotic levels. Esin Yuluğ, Sibel Türedi, Ahmet Alver, Süleyman Türedi, and Cemil Kahraman Copyright © 2013 Esin Yuluğ et al. All rights reserved. Clinical Detoxification: Elimination of Persistent Toxicants from the Human Body Thu, 06 Jun 2013 11:30:10 +0000 Stephen J. Genuis, Margaret E. Sears, Gerry Schwalfenberg, Janette Hope, and Robin Bernhoft Copyright © 2013 Stephen J. Genuis et al. All rights reserved. Cadmium Toxicity and Treatment Mon, 03 Jun 2013 15:45:10 +0000 Cadmium is a heavy metal of considerable toxicity with destructive impact on most organ systems. It is widely distributed in humans, the chief sources of contamination being cigarette smoke, welding, and contaminated food and beverages. Toxic impacts are discussed and appear to be proportional to body burden of cadmium. Detoxification of cadmium with EDTA and other chelators is possible and has been shown to be therapeutically beneficial in humans and animals when done using established protocols. Robin A. Bernhoft Copyright © 2013 Robin A. Bernhoft. All rights reserved. A Review of the Mechanism of Injury and Treatment Approaches for Illness Resulting from Exposure to Water-Damaged Buildings, Mold, and Mycotoxins Thu, 18 Apr 2013 13:51:03 +0000 Physicians are increasingly being asked to diagnose and treat people made ill by exposure to water-damaged environments, mold, and mycotoxins. In addition to avoidance of further exposure to these environments and to items contaminated by these environments, a number of approaches have been used to help persons affected by exposure to restore their health. Illness results from a combination of factors present in water-damaged indoor environments including, mold spores and hyphal fragments, mycotoxins, bacteria, bacterial endotoxins, and cell wall components as well as other factors. Mechanisms of illness include inflammation, oxidative stress, toxicity, infection, allergy, and irritant effects of exposure. This paper reviews the scientific literature as it relates to commonly used treatments such as glutathione, antioxidants, antifungals, and sequestering agents such as Cholestyramine, charcoal, clay and chlorella, antioxidants, probiotics, and induced sweating. Janette Hope Copyright © 2013 Janette Hope. All rights reserved. Chelation: Harnessing and Enhancing Heavy Metal Detoxification—A Review Thu, 18 Apr 2013 13:44:58 +0000 Toxic metals such as arsenic, cadmium, lead, and mercury are ubiquitous, have no beneficial role in human homeostasis, and contribute to noncommunicable chronic diseases. While novel drug targets for chronic disease are eagerly sought, potentially helpful agents that aid in detoxification of toxic elements, chelators, have largely been restricted to overt acute poisoning. Chelation, that is multiple coordination bonds between organic molecules and metals, is very common in the body and at the heart of enzymes with a metal cofactor such as copper or zinc. Peptides glutathione and metallothionein chelate both essential and toxic elements as they are sequestered, transported, and excreted. Enhancing natural chelation detoxification pathways, as well as use of pharmaceutical chelators against heavy metals are reviewed. Historical adverse outcomes with chelators, lessons learned in the art of using them, and successes using chelation to ameliorate renal, cardiovascular, and neurological conditions highlight the need for renewed attention to simple, safe, inexpensive interventions that offer potential to stem the tide of debilitating, expensive chronic disease. Margaret E. Sears Copyright © 2013 Margaret E. Sears. All rights reserved. Toxicological Profile of Chlorophenols and Their Derivatives in the Environment: The Public Health Perspective Wed, 03 Apr 2013 17:32:13 +0000 Chlorophenol compounds and their derivatives are ubiquitous contaminants in the environment. These compounds are used as intermediates in manufacturing agricultural chemicals, pharmaceuticals, biocides, and dyes. Chlorophenols gets into the environment from a variety of sources such as industrial waste, pesticides, and insecticides, or by degradation of complex chlorinated hydrocarbons. Thermal and chemical degradation of chlorophenols leads to the formation of harmful substances which constitute public health problems. These compounds may cause histopathological alterations, genotoxicity, mutagenicity, and carcinogenicity amongst other abnormalities in humans and animals. Furthermore, the recalcitrant nature of chlorophenolic compounds to degradation constitutes an environmental nuisance, and a good understanding of the fate and transport of these compounds and their derivatives is needed for a clearer view of the associated risks and mechanisms of pathogenicity to humans and animals. This review looks at chlorophenols and their derivatives, explores current research on their effects on public health, and proffers measures for mitigation. Etinosa O. Igbinosa, Emmanuel E. Odjadjare, Vincent N. Chigor, Isoken H. Igbinosa, Alexander O. Emoghene, Fredrick O. Ekhaise, Nicholas O. Igiehon, and Omoruyi G. Idemudia Copyright © 2013 Etinosa O. Igbinosa et al. All rights reserved. An Assessment of the Intestinal Lumen as a Site for Intervention in Reducing Body Burdens of Organochlorine Compounds Thu, 07 Feb 2013 15:21:47 +0000 Many individuals maintain a persistent body burden of organochlorine compounds (OCs) as well as other lipophilic compounds, largely as a result of airborne and dietary exposures. Ingested OCs are typically absorbed from the small intestine along with dietary lipids. Once in the body, stored OCs can mobilize from adipose tissue storage sites and, along with circulating OCs, are delivered into the small intestine via hepatic processing and biliary transport. Retained OCs are also transported into both the large and small intestinal lumen via non-biliary mechanisms involving both secretion and desquamation from enterocytes. OCs and some other toxicants can be reabsorbed from the intestine, however, they take part in enterohepatic circulation(EHC). While dietary fat facilitates the absorption of OCs from the small intestine, it has little effect on OCs within the large intestine. Non-absorbable dietary fats and fat absorption inhibitors, however, can reduce the re-absorption of OCs and other lipophiles involved in EHC and may enhance the secretion of these compounds into the large intestine—thereby hastening their elimination. Clinical studies are currently underway to determine the efficacy of using non-absorbable fats and inhibitors of fat absorption in facilitating the elimination of persistent body burdens of OCs and other lipophilic human contaminants. Ronald J. Jandacek and Stephen J. Genuis Copyright © 2013 Ronald J. Jandacek and Stephen J. Genuis. All rights reserved. Human Elimination of Phthalate Compounds: Blood, Urine, and Sweat (BUS) Study Thu, 01 Nov 2012 09:51:22 +0000 Background. Individual members of the phthalate family of chemical compounds are components of innumerable everyday consumer products, resulting in a high exposure scenario for some individuals and population groups. Multiple epidemiological studies have demonstrated statistically significant exposure-disease relationships involving phthalates and toxicological studies have shown estrogenic effects in vitro. Data is lacking in the medical literature, however, on effective means to facilitate phthalate excretion. Methods. Blood, urine, and sweat were collected from 20 individuals (10 healthy participants and 10 participants with assorted health problems) and analyzed for parent phthalate compounds as well as phthalate metabolites using high performance liquid chromatography-tandem mass spectrometry. Results. Some parent phthalates as well as their metabolites were excreted into sweat. All patients had MEHP (mono(2-ethylhexyl) phthalate) in their blood, sweat, and urine samples, suggesting widespread phthalate exposure. In several individuals, DEHP (di (2-ethylhexl) phthalate) was found in sweat but not in serum, suggesting the possibility of phthalate retention and bioaccumulation. On average, MEHP concentration in sweat was more than twice as high as urine levels. Conclusions. Induced perspiration may be useful to facilitate elimination of some potentially toxic phthalate compounds including DEHP and MEHP. Sweat analysis may be helpful in establishing the existence of accrued DEHP in the human body. Stephen J. Genuis, Sanjay Beesoon, Rebecca A. Lobo, and Detlef Birkholz Copyright © 2012 Stephen J. Genuis et al. All rights reserved. Antibiotic, Pesticide, and Microbial Contaminants of Honey: Human Health Hazards Sun, 14 Oct 2012 19:46:35 +0000 Agricultural contamination with pesticides and antibiotics is a challenging problem that needs to be fully addressed. Bee products, such as honey, are widely consumed as food and medicine and their contamination may carry serious health hazards. Honey and other bee products are polluted by pesticides, heavy metals, bacteria and radioactive materials. Pesticide residues cause genetic mutations and cellular degradation and presence of antibiotics might increase resistant human or animal's pathogens. Many cases of infant botulisms have been attributed to contaminated honey. Honey may be very toxic when produced from certain plants. Ingestion of honey without knowing its source and safety might be problematic. Honey should be labeled to explore its origin, composition, and clear statement that it is free from contaminants. Honey that is not subjected for analysis and sterilization should not be used in infants, and should not be applied to wounds or used for medicinal purposes. This article reviews the extent and health impact of honey contamination and stresses on the introduction of a strict monitoring system and validation of acceptable minimal concentrations of pollutants or identifying maximum residue limits for bee products, in particular, honey. Noori Al-Waili, Khelod Salom, Ahmed Al-Ghamdi, and Mohammad Javed Ansari Copyright © 2012 Noori Al-Waili et al. All rights reserved. Deriving Freshwater Quality Criteria for Iron, Lead, Nickel, and Zinc for Protection of Aquatic Life in Malaysia Thu, 02 Aug 2012 10:56:48 +0000 Freshwater quality criteria for iron (Fe), lead (Pb), nickel (Ni), and zinc (Zn) were developed with particular reference to aquatic biota in Malaysia, and based on USEPA’s guidelines. Acute toxicity tests were performed on eight different freshwater domestic species in Malaysia which were Macrobrachium lanchesteri (prawn), two fish: Poecilia reticulata and Rasbora sumatrana, Melanoides tuberculata (snail), Stenocypris major (ostracod), Chironomus javanus (midge larvae), Nais elinguis (annelid), and Duttaphrynus melanostictus (tadpole) to determine 96 h LC50 values for Fe, Pb, Ni, and Zn. The final acute value (FAV) for Fe, Pb, Ni, and Zn were 74.5, 17.0, 165, and 304.9 μg L−1, respectively. Using an estimated acute-to-chronic ratio (ACR) of 8.3, the value for final chronic value (FCV) was derived. Based on FAV and FCV, a criterion maximum concentration (CMC) and a criterion continuous concentration (CCC) for Fe, Pb, Ni, and Zn that are 37.2, 8.5, 82.5, and 152.4 μg L−1 and 9.0, 2.0, 19.9, and 36.7 μg L−1, respectively, were derived. The results of this study provide useful data for deriving national or local water quality criteria for Fe, Pb, Ni, and Zn based on aquatic biota in Malaysia. Based on LC50 values, this study indicated that N. elinguis, M. lanchesteri, N. elinguis, and R. sumatrana were the most sensitive to Fe, Pb, Ni, and Zn, respectively. M. Shuhaimi-Othman, Y. Nadzifah, R. Nur-Amalina, and N. S. Umirah Copyright © 2012 M. Shuhaimi-Othman et al. All rights reserved. The Effects of Subchronic Exposure to Metribuzin on Danio rerio Mon, 18 Jun 2012 10:27:34 +0000 The aim of this study was to assess the impact of metribuzin in surface waters on fish under experimental conditions. The effects of subchronic exposure to metribuzin on fish growth and the development of histopathological changes in selected organs (gill, kidney, liver) and on activity of some biochemical markers (CYP450, EROD) in Danio rerio were investigated during a 28-day toxicity test. Juvenile growth tests were performed on D. rerio according to OECD method number 215. Fish at an initial age of 30 days were exposed to a range of metribuzin concentrations (1.5, 5, 16, 33, and 53 mg L−1). Exposure to metribuzin at 53 mg L−1 was associated with increased mortality. Negative effects with regard to total body weight, length, and the inhibition of specific growth rate were induced at concentrations of 33 and 53 mg L−1. Histopathological examination revealed pathological lesions in the liver in pesticide-exposed fish only at the highest concentration of 53 mg L−1 of metribuzin. Based on the results of growth rate, biochemical markers (CYP450, EROD), and histopathological examination, the lowest observed effect concentration (LOEC) value was 33 mg L−1 and the no observed effect concentration (NOEC) value was 16 mg L−1. Lucie Plhalova, Stanislava Stepanova, Eva Praskova, Lucie Chromcova, Lenka Zelnickova, Lenka Divisova, Misa Skoric, Vladimira Pistekova, Iveta Bedanova, and Zdenka Svobodova Copyright © 2012 Lucie Plhalova et al. All rights reserved. Assessment of Coumarin Levels in Ground Cinnamon Available in the Czech Retail Market Mon, 18 Jun 2012 10:12:52 +0000 The objective of this study was to determine the coumarin content of ground cinnamon purchased from retail markets in the Czech Republic. No sample was labelled with information on the botanical source, but, in some cases, the countries of origin were specified. For comparison, a single cinnamon sample imported directly from a plantation in Sri Lanka that came from Cinnamomum verum was analyzed. Results from 60 ground cinnamon samples comprising twelve brands confirmed a high content of coumarin, with mean levels ranging from 2 650 to 7 017 mg · kg−1. The high coumarin content confirmed that these cinnamon samples obtained from cassia cinnamon were in contrast to the sample from Sri Lanka, which was coumarin-free. Jana Blahová and Zdeňka Svobodová Copyright © 2012 Jana Blahová and Zdeňka Svobodová. All rights reserved. Effect of Magnesium Supplementation on the Distribution Patterns of Zinc, Copper, and Magnesium in Rabbits Exposed to Prolonged Cadmium Intoxication Mon, 04 Jun 2012 15:35:50 +0000 The present study is designed to investigate whether magnesium (Mg) supplementation may prevent Cd-induced alterations in zinc (Zn), copper (Cu), and magnesium (Mg) status in rabbits. For this purpose, the concentrations of Zn, Cu, and Mg were estimated in blood, urine, and organs (brain, heart, lungs, liver, kidney, spleen, pancreas, skeletal muscle, and bone) of rabbits given Cd (10 mg/kg b.w.) and rabbits cotreated with Mg (40 mg/kg b.w.) orally, as aqueous solutions of Cd chloride and Mg acetate every day for 4 weeks. Samples were mineralized with conc. HNO3 and HClO4 (4:1) and metals concentrations were determined by atomic absorption spectrophotometry (AAS). Magnesium supplementation succeeded to overcome Cd-induced disbalance of investigated bioelements. Beneficial effects of Mg were observed on Zn levels in blood and urine, on Cu levels in urine, and on Mg levels in blood. Magnesium pretreatment also managed to counteract or reduce all Cd-induced changes in levels of Cu and Mg in organs, while it did not exert this effect on Zn levels. These findings suggest that enhanced dietary Mg intake during Cd exposure can have at least partly beneficial effect on Cd-induced alterations in homeostasis of zinc, copper, and magnesium. Zorica Bulat, Danijela Ðukic-Cosic, Biljana Antonijevic, Petar Bulat, Dragana Vujanovic, Aleksandra Buha, and Vesna Matovic Copyright © 2012 Zorica Bulat et al. All rights reserved. The Effects of Subchronic Exposure to Terbuthylazine on Early Developmental Stages of Common Carp Wed, 02 May 2012 14:36:04 +0000 The aim of this study was to assess the impact of terbuthylazine in surface waters on fish under experimental conditions. Subchronic toxic effects on embryos and larvae of common carp (Cyprinus carpio) were investigated during a 30-day toxicity test. The exposure to terbuthylazin showed no effect on mortality, but significant differences (𝑃<0.0001) were revealed on weight and growth parameters at concentrations of 520 and 820 μg/L. The inhibition of specific growth rate at concentrations of 520 and 820 μg/L was 14% compared to the control group. No significant negative effects on total body length and body weight were observed at lower concentrations (0.9 and 160 μg/L). The concentrations 520 and 820 μg/L were associated with a delay in development compared to other experimental groups and controls. On the basis of weight and growth rate evaluation and determination of developmental stages, the No Observed Effect Concentration (NOEC) of terbuthylazine was estimated at 160 μg/L and the Lowest Observed Effect Concentration (LOEC) was 520 μg/L. According to these results, the reported environmental concentration of terbuthylazine in Czech rivers does not impact growth, development, morphology, or histology of carp embryos and larvae. Stanislava Štepánová, Lucie Plhalová, Petra Doleželová, Miroslav Prokeš, Petr Maršálek, Miša Škoric, and Zdenka Svobodová Copyright © 2012 Stanislava Štěpánová et al. All rights reserved. Are Fish and Standardized FETAX Assays Protective Enough for Amphibians? A Case Study on Xenopus laevis Larvae Assay with Biologically Active Substances Present in Livestock Wastes Tue, 01 May 2012 19:08:10 +0000 Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC50) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians. Federica Martini, José V. Tarazona, and M. Victoria Pablos Copyright © 2012 Federica Martini et al. All rights reserved. General and Electrophysiological Toxic Effects of Manganese in Rats following Subacute Administration in Dissolved and Nanoparticle Form Tue, 01 May 2012 15:53:39 +0000 In an attempt to model occupational and environmental Mn exposures and their possible interaction, young male Wistar rats were exposed to Mn by oral administration in dissolved form (MnCl2·4H2O, 14.84 and 59.36 mg/kg b.w.) and by intratracheal application of MnO2 nanoparticles (2.63 mg/kg b.w.). After 3 and 6 weeks oral, or 3 weeks oral plus 3 weeks intratracheal, exposure, general toxicological, and electrophysiological tests were done. Body weight gain was significantly reduced after 6 and 3 plus 3 weeks exposure, but the effect of the latter on the pace of weight gain was stronger. Organ weights signalized systemic stress and effect on lungs. Changes in evoked electrophysiological responses (cortical sensory evoked potential and nerve action potential) indicated that the 3 plus 3 weeks combined exposure caused equal or higher changes in the latency of these responses than 6 weeks of exposure, although the calculated summed Mn dose in the former case was lower. The results showed the importance of the physicochemical form of Mn in determining the toxic outcome, and suggested that neurofunctional markers of Mn action may indicate the human health effect better than conventional blood Mn measurement. Edina Horváth, Zsuzsanna Máté, Szabolcs Takács, Péter Pusztai, András Sápi, Zoltán Kónya, László Nagymajtényi, and András Papp Copyright © 2012 Edina Horváth et al. All rights reserved. Impact of Cypermethrin on Fingerlings of Common Edible Carp (Labeo rohita) Mon, 30 Apr 2012 14:41:00 +0000 Laboratory evaluations were made to assess the toxicological and biochemical effect of cypermethrin on fingerlings of common edible freshwater culture carp (Labeo rohita). There was a significant negative (𝑃<0.05) correlation observed between effective doses of cypermethrin and exposure periods; that is, LC50 values decreased from 0.323 μg/L (6 h) to  > 0.278 μg/L (12 h),  > 0.240 μg/L (18 h) and >0.205 μg/L (24 h). Exposure to sublethal doses of cypermethrin for 24 h and 96 h exposure period caused significant (𝑃<0.05) time- and dose-dependent alterations in total protein, total free amino acids, nucleic acids, glycogen, pyruvate, and lactate level and in the activity of enzyme protease, alanine aminotransferase, aspartate aminotransferase, acid phosphatases, alkaline phosphatases, acetylcholinesterase, and cytochrome oxidase in liver and muscle tissues of fish. Thus, cypermethrin has potent piscicidal activity against fingerlings of fish Labeo rohita and adversely affects their behavioural patterns, shifting aerobic pathway of fish respiration towards anaerobic pathway and also inhibiting energy production by suppressing ATP synthesis. Sudhanshu Tiwari, Richa Tiwari, and Ajay Singh Copyright © 2012 Sudhanshu Tiwari et al. All rights reserved. Salicylic Acid Attenuates Gentamicin-Induced Nephrotoxicity in Rats Mon, 30 Apr 2012 13:10:43 +0000 Gentamicin (GM) is a widely used antibiotic against serious and life-threatening infections, but its usefulness is limited by the development of nephrotoxicity. The present study was designed to determine the protective effect of salicylic acid (SA) in gentamicin-induced nephrotoxicity in rats. Quantitative evaluation of gentamicin-induced structural alterations and degree of functional alterations in the kidneys were performed by histopathological and biochemical analyses in order to determine potential beneficial effects of SA coadministration with gentamicin. Gentamicin was observed to cause a severe nephrotoxicity which was evidenced by an elevation of serum urea and creatinine levels. The significant increases in malondialdehyde (MDA) levels and protein carbonyl groups indicated that GM-induced tissue injury was mediated through oxidative reactions. On the other hand, simultaneous SA administration protected kidney tissue against the oxidative damage and the nephrotoxic effect caused by GM treatment. Exposure to GM caused necrosis of tubular epithelial cells. Necrosis of tubules was found to be prevented by SA pretreatment. The results from our study indicate that SA supplement attenuates oxidative-stress associated renal injury by reducing oxygen free radicals and lipid peroxidation in gentamicin-treated rats. Pavle Randjelovic, Slavimir Veljkovic, Nenad Stojiljkovic, Ljubinka Jankovic-Velickovic, Dusan Sokolovic, Milan Stoiljkovic, and Ivan Ilic Copyright © 2012 Pavle Randjelovic et al. All rights reserved. The Growth of Monoraphidium sp. and Scenedesmus sp. Cells in the Presence of Thorium Sun, 29 Apr 2012 12:03:59 +0000 Toxicity of thorium by Monoraphidium sp. and Scenedesmus sp. was studied. Microalgal cultures were inoculated in ASM-1 medium in presence and absence of thorium. Its effect was monitored by direct counting on Fuchs-Rosenthal chamber and with software. The toxicity of thorium over the species was observed for concentrations over 50.0 mg/L. After 30 days, Monoraphidium cells decreased their concentration from 4.23×106 to 4.27×105 and 8.57×105 cells/mL, in the presence of 50.0 and 100.0 mg/L of thorium, respectively. Scenedesmus sp. cells were more resistant to thorium: for an initial cell concentration of 7.65×104 cells/mL it was observed a change to 5.25×105 and 5.12×105 cells/mL, in the presence of thorium at 50.0 and 100.0 mg/L, respectively. This is an indication that low concentrations of the radionuclide favored the growth, and that Scenedesmus cells are more resistant to thorium than Monoraphidium cells. The software used for comparison with direct count method proved to be useful for the improvement of accuracy of the results obtained, a decrease in the uncertainty and allowed recording of the data. The presence of thorium suggests that low concentrations have a positive effect on the growth, due to the presence of the nitrate, indicating its potential for ecotoxicological studies. Juliana Cristina de Queiroz, Ana Cristina de Melo Ferreira, and Antonio Carlos Augusto da Costa Copyright © 2012 Juliana Cristina de Queiroz et al. All rights reserved. Acute and Chronic Toxicity of Soluble Fractions of Industrial Solid Wastes on Daphnia magna and Vibrio fischeri Sun, 22 Apr 2012 07:59:24 +0000 Industrial wastes may produce leachates that can contaminate the aquatic ecosystem. Toxicity testing in acute and chronic levels is essential to assess environmental risks from the soluble fractions of these wastes, since only chemical analysis may not be adequate to classify the hazard of an industrial waste. In this study, ten samples of solid wastes from textile, metal-mechanic, and pulp and paper industries were analyzed by acute and chronic toxicity tests with Daphnia magna and Vibrio fischeri. A metal-mechanic waste (sample MM3) induced the highest toxicity level to Daphnia magna(CE50,48h=2.21%). A textile waste induced the highest toxicity level to Vibrio fischeri (sample TX2, CE50,30min=12.08%). All samples of pulp and paper wastes, and a textile waste (sample TX2) induced chronic effects on reproduction, length, and longevity of Daphnia magna. These results could serve as an alert about the environmental risks of an inadequate waste classification method. Letícia Flohr, Armando Borges de Castilhos Júnior, and William Gerson Matias Copyright © 2012 Letícia Flohr et al. All rights reserved. Efficacy and Safety of Immunotherapy with Interferon-Gamma in the Management of Chronic Sulfur Mustard-Induced Cutaneous Complications: Comparison with Topical Betamethasone 1% Mon, 12 Mar 2012 14:56:11 +0000 The present trial investigated the efficacy of immunotherapy with interferon-gamma (IFN-γ) in the treatment of sulfur mustard (SM)-induced chronic skin complications. Forty subjects who were suffering from chronic skin complications of SM and were diagnosed to have severe atopic dermatitis, were assigned to IFN-γ (50 μg/m2) subcutaneously three times per week (𝑛=20) or betamethasone valerate topical cream 0.1% (𝑛=20) every night for 30 days. Extent and intensity of cutaneous complications was evaluated using scoring atopic dermatitis (SCORAD) index, and quality of life using dermatology life quality index (DLQI) at baseline and at the end of trial. SCORAD-A and SCORAD-B scores were significantly decreased in both IFN-γ and betamethasone. However, SCORAD-C score was decreased only in the IFN-γ group. There were significant reductions in overall as well as objective SCORAD scores in both groups. As for the magnitude of changes, treatment with IFN-γ was associated with greater reductions in overall, objective and segmented SCORAD scores compared to betamethasone. DLQI reduction was found to be significantly greater in the IFN-γ group. Promising improvements in quality life and clinical symptoms that was observed in the present study suggest the application of IFN-γ as an effective therapy for the management of SM-induced chronic skin complications. Yunes Panahi, Amirhossein Sahebkar, Seyyed Masoud Davoudi, Mojtaba Amiri, and Fatemeh Beiraghdar Copyright © 2012 Yunes Panahi et al. All rights reserved. Levels of Heavy Metals in Popular Cigarette Brands and Exposure to These Metals via Smoking Mon, 12 Mar 2012 13:45:22 +0000 The levels of selected heavy metals in popular cigarette brands sold and/or produced in Saudi Arabia were determined by graphite furnace-atomic absorption spectrometry (GFAAS). Average concentrations of Cadmium and Lead in different cigarette brands were 1.81 and 2.46 μg g−1 (dry weight), respectively. The results obtained in this study estimate the average quantity of Cd inhaled from smoking one packet of 20 cigarettes to be in the range of 0.22–0.78 μg. Results suggest that the quantity of Pb inhaled of smoking one packet of 20 cigarettes is estimated to be 0.97–2.64 μg. The concentrations of Cd and Pb in cigarettes were significantly different between cigarette brands tested. The results of the present study were compared with those of other regional and international studies. Muhammad Waqar Ashraf Copyright © 2012 Muhammad Waqar Ashraf. All rights reserved. Polychlorinated Biphenyls-Induced Oxidative Stress on Rat Hippocampus: A Neuroprotective Role of Quercetin Wed, 04 Jan 2012 10:58:11 +0000 Present study is aimed to evaluate the ameliorative role of quercetin on PCBs-induced oxidative stress in hippocampus of Wistar rats. Group I rats received vehicle (corn oil) intraperitoneally (i.p); Group II received quercetin 50 mg/kg bwt/day (gavage); Group III received PCB 2 mg/kg bwt/day (i.p); Group IV received PCB (i.p) and simultaneously quercetin through gavage. After 30 days, rats were euthanized and hippocampus was dissected from each rat brain. Oxidative stress was assessed by determining the levels of H2O2, LPO, Pcc, and alteration in the functional markers such as CK, AchE, and ATPases activities in the hippocampus of control and experimental animals. A significant increase in the levels of stress markers and decrease in level of functional markers were observed in PCBs-treated rats. Moreover DNA fragmentation and histological studies were ascertained to confirm PCBs toxicity. In conclusion, quercetin shows a protective role against PCBs-induced oxidative damage in rat hippocampus. Kandaswamy Selvakumar, Senthamilselvan Bavithra, Gunasekaran Krishnamoorthy, Prabhu Venkataraman, and Jagadeesan Arunakaran Copyright © 2012 Kandaswamy Selvakumar et al. All rights reserved. The Inflammatory Process in Response to Nanoparticles Thu, 15 Dec 2011 00:00:00 +0000 David M. Goncalves, Rafael de Liz, and Denis Girard Copyright © 2011 David M. Goncalves et al. All rights reserved. Biochemical and Ultrastructural Changes in the Hepatopancreas of Bellamya aeruginosa (Gastropoda) Fed with Toxic Cyanobacteria Tue, 01 Nov 2011 00:00:00 +0000 This study was conducted to investigate ultrastructural alterations and biochemical responses in the hepatopancreas of the freshwater snail Bellamya aeruginosa after exposure to two treatments: toxic cyanobacterium (Microcystis aeruginosa) and toxic cyanobacterial cells mixed with a non-toxic green alga (Scendesmus quadricauda) for a period of 15 days of intoxication, followed by a 15-day detoxification period. The toxic algal suspension induced a very pronounced increase of the activities of acid phosphatases, alkaline phosphatases and glutathione S-transferases (ACP, ALP and GST) in the liver at the later stage of intoxication. During the depuration, enzymatic activity tended to return to the levels close to those in the control. The activity of GST displayed the most pronounced response among different algal suspensions. Severe cytoplasmic vacuolization, condensation and deformation of nucleus, dilation and myeloid-like in mitochondria, disruption of rough endoplasmic reticulum, proliferation of lysosome, telolysosomes and apoptotic body were observed in the tissues. All cellular organelles began recovery after the snails were transferred to the S. quadricauda. The occurrence of a large amount of activated lysosomes and heterolysosomes and augment in activity of detoxification enzyme GST might be an adaptive mechanism to eliminate or lessen cell damage caused by hepatotoxicity to B. aeruginosa. Jinyong Zhu, Kaihong Lu, Chunjing Zhang, Jingjing Liang, and Zhiyong Hu Copyright © 2011 Jinyong Zhu et al. All rights reserved. Activation of Neutrophils by Nanoparticles Mon, 24 Oct 2011 00:00:00 +0000 The use of nanoparticles (NPs) has increased in the past few years in various fields, including defence, aerospace, electronics, biology, medicine, and so forth. and in applications such as diagnostic technology, bioimaging, and drug/gene delivery. Thus, human exposure to NPs and nanomaterials is unavoidable and will certainly expand in the future resulting in a growing interest in nanotoxicology, the study of toxicity of nanomaterials. A number of studies have reported the effects of NPs in respect to pulmonary inflammation by investigating in vitro activation of pulmonary cells with NPs and in vivo in a variety of models in which neutrophils appear to be the predominant leukocyte cell type in lungs and in bronchoalveolar lavages following inhalation or intratracheal instillation of NPs. Despite the fact that several studies have reported an increased number of neutrophils, the literature dealing with the direct activation of neutrophils by a given NP is poorly documented. This paper will summarize the current literature in this latter area of research and will end with a perspective view in which our laboratory will be involved in the following years. David M. Goncalves, Rafael de Liz, and Denis Girard Copyright © 2011 David M. Goncalves et al. All rights reserved. Nanoexposure, Unusual Diseases, and New Health and Safety Concerns Thu, 20 Oct 2011 00:00:00 +0000 Accumulating studies in animals have shown that nanoparticles could cause unusual rapid lung injury and extrapulmonary toxicity. Whether exposure of workers to nanoparticles may result in some unexpected damage as seen in animals is still a big concern. We previously reported findings regarding a group of patients exposed to nanoparticles and presenting with an unusual disease. The reported disease was characterized by bilateral chest fluid, pulmonary fibrosis, pleural granuloma, and multiorgan damage and was highly associated with the nanoparticle exposure. To strengthen this association, further information on exposure and the disease was collected and discussed. Our studies show that some kinds of nanomaterials, such as silica nanoparticles and nanosilicates, may be very toxic and even fatal to occupational workers exposed to them without any effective personal protective equipment. More research and collaborative efforts on nanosafety are required in order to prevent and minimize the potential hazards of nanomaterials to humans and the environment. Yuguo Song and Shichuan Tang Copyright © 2011 Yuguo Song and Shichuan Tang. All rights reserved. Evaluating Side Effects of Human Exposure to Noxious Agents Mon, 01 Jan 1900 00:00:00 +0000 Demetrios P. Matthopoulos Copyright © 2006 Demetrios P. Matthopoulos. All rights reserved. Distribution, Levels, and Risk Assessment of Polycyclic Aromatic Hydrocarbons (PAHs) in Some Water Bodies along the Coastal Belt of Ghana Mon, 01 Jan 1900 00:00:00 +0000 The levels and distribution of 24 polycyclic aromatic hydrocarbons (PAHs) were determined in six water bodies along the coastal belt of Ghana using gas chromatography with flame-ionization detection (GC/FID). The average total PAHs recorded are from the Pra estuary, 6.3 μg/L; Benya lagoon, 7.5 μg/L; Sakumono lagoon, 10.1 μg/L; lower Volta estuary, 26.3 μg/L; Keta lagoon, 10.6 μg/L; and Narkwa lagoon, 16.1 μg/L.The 12 PAHs that were well distributed in all the coastal waters analyzed include naphthalene, pyrene, fluorene, 2-methylnaphthalene, 2,6-dimethylnaphthalene, acephnaphthalene, acephnaphthene, 1-methylphenanthrene, 2,3,5-trimethylnaphthalene, chrysene, biphenyl, and phenanthrene. The presence of benzo(b)fluoranthene, benzo(a)anthracene, and benzo(j,k)fluoranthene in some of the water bodies is a source of concern as they have been classified by the International Agency for Research on Cancer (IARC) and U.S. Environmental Protection Agency (EPA) as probable human carcinogens. These water bodies are used for fishing and for some domestic purposes by the people living around them, thereby exposing them to some dangers and the risk of getting cancer. The human health cancer risk assessment carried out also indicates that there is the possibility of some users of the water bodies getting cancer in their lifetime. David Kofi Essumang Copyright © 2010 David Kofi Essumang. All rights reserved. Toxicity of Single and Mixed Contaminants in Seawater Measured with Acute Toxicity Bioassays Mon, 01 Jan 1900 00:00:00 +0000 Different types of organic pollutants commonly detected in seawater have been evaluated by acute toxicity bioassays. Vibrio fischeri, Daphnia magna, and Selenastrum capricornotum were selected to test toxic effects of individual compounds and mixtures of these compounds, obtaining EC50 values in the range of 0.001 to 28.9 mg/l. In the case of mixtures, synergistic toxic responses were seen for a clear majority of the cases (>60%). Mixtures containing methyl-tertiary-butyl ether (MTBE) exhibit accelerated processes that result in a change in concentration required to produce a toxic effect; for example, in the case of mixtures containing MTBE and Diuron and Dichlofluanid. A.R. Fernandez-Alba, L. Piedra, M. Mezcua, and M.D. Hernando Copyright © 2002 A.R. Fernandez-Alba et al. All rights reserved. Genotoxicity and Mutagenicity of Suspended Particulate Matter of River Water and Waste Water Samples Mon, 01 Jan 1900 00:00:00 +0000 Suspended particulate matter of samples of river water and waste water treatment plants was tested for genotoxicity and mutagenicity using the standardized umu assay and two versions of the Ames microsuspension assay. The study tries to determine the entire DNA-damaging potential of the water samples and the distribution of DNA-damaging substances among the liquid phase and solid phase. Responsiveness and sensitivity of the bioassays are compared. Georg Reifferscheid and Britta v. Oepen Copyright © 2002 Georg Reifferscheid and Britta v. Oepen. All rights reserved. Toxicity of Flare and Crude Hydrocarbon Mixtures Mon, 01 Jan 1900 00:00:00 +0000 The toxicity of whole, saturate, and aromatic hydrocarbon mixtures from flare pit and crude oil sources were evaluated using Lumbricus terrestris. Body burden analysis was used to analyze the intrinsic toxicity of the six hydrocarbon mixtures. The major fractions of the whole mixtures, the saturate, and aromatic fractions had different intrinsic toxicities; the aromatics were more toxic than the saturates. The toxicity of the saturate and aromatic fractions also differed between the mixtures. The flare saturate mixture was more toxic than the crude saturate mixture, while the crude aromatic mixture was more toxic than the flare aromatic mixture. The most dramatic difference in toxicity of the two sources was between the flare whole and crude whole mixtures. The crude whole mixture was very toxic; the toxicity of this mixture reflected the toxicity of the crude aromatic fraction. However, the flare whole mixture was not toxic, due to a lack of partitioning from the whole mixture into the lipid membrane of the exposed worms. This lack of partitioning appears to be related to the relatively high concentrations of asphaltenes and polar compounds in the flare pit whole mixture. Sonja V. Cook, Angus Chu, and Ron Goodman Copyright © 2002 Sonja V. Cook et al. All rights reserved. Parameterization of the Stomatal Component of the DO3SE Model for Mediterranean Evergreen Broadleaf Species Mon, 01 Jan 1900 00:00:00 +0000 An ozone (O3) deposition model (DO3SE) is currently used in Europe to define the areas where O3 concentrations lead to absorbed O3 doses that exceed the flux-based critical levels above which phytotoxic effects would be likely recorded. This mapping exercise relies mostly on the accurate estimation of O3 flux through plant stomata. However, the present parameterization of the modulation of stomatal conductance (gs) behavior by different environmental variables needs further adjustment if O3 phytotoxicity is to be assessed accurately at regional or continental scales. A new parameterization of the model is proposed for Holm oak (Quercus ilex), a tree species that has been selected as a surrogate for all Mediterranean evergreen broadleaf species. This parameterization was based on a literature review, and was calibrated and validated using experimentally measured data of gs and several atmospheric and soil parameters recorded at three sites of the Iberian Peninsula experiencing long summer drought, and very cold and dry winter air (El Pardo and Miraflores) or milder conditions (Tietar). A fairly good agreement was found between modeled and measured data (R2 = 0.64) at Tietar. However, a reasonable performance (R2 = 0.47–0.62) of the model was only achieved at the most continental sites when gs and soil moisture deficit relationships were considered. The influence of root depth on gs estimation is discussed and recommendations are made to build up separate parameterizations for continental and marine-influenced Holm oak sites in the future. Roccío Alonso, Susana Elvira, María J. Sanz, Lisa Emberson, and Benjamín S. Gimeno Copyright © 2007 Roccio Alonso et al. All rights reserved. Investigation of Arsenic-Stressed Yeast (Saccharomyces cerevisiae) as a Bioassay in Homeopathic Basic Research Mon, 01 Jan 1900 00:00:00 +0000 This study investigated the response of arsenic-stressed yeast (Saccharomyces cerevisiae) towards homeopathically potentized Arsenicum album, a duckweed nosode, and gibberellic acid. The three test substances were applied in five potency levels (17x, 18x, 24x, 28x, 30x) and compared to controls (unsuccussed and succussed water) with respect to influencing specific growth parameters. Five independent experiments were evaluated for each test substance. Additionally, five water control experiments were analyzed to investigate the stability of the experimental setup (systematic negative control experiments). All experiments were randomized and blinded. Yeast grew in microplates over a period of 38 h in either potentized substances or water controls with 250 mg/l arsenic(V) added over the entire cultivation period. Yeast's growth kinetics (slope, Et50, and yield) were measured photometrically. The test system exhibited a low coefficient of variation (slope 1.2%, Et50 0.3%, yield 2.7%). Succussed water did not induce any significant differences compared to unsuccussed water. Data from the control and treatment groups were both pooled to increase statistical power. In this study with yeast, no significant effects were found for any outcome parameter or any homeopathic treatment. Since in parallel experiments arsenic-stressed duckweed showed highly significant effects after application of potentized Arsenicum album and duckweed nosode preparations from the same batch as used in the present study, some specific properties of this experimental setup with yeast must be responsible for the lacking response. Tim Jäger, Claudia Scherr, Ursula Wolf, Meinhard Simon, Peter Heusser, and Stephan Baumgartner Copyright © 2011 Tim Jäger et al. All rights reserved. Impacts of Extraction Methods in the Rapid Determination of Atrazine Residues in Foods using Supercritical Fluid Chromatography and Enzyme-Linked Immunosorbent Assay: Microwave Solvent vs. Supercritical Fluid Extractions Mon, 01 Jan 1900 00:00:00 +0000 It is an accepted fact that many food products that we eat today have the possibility of being contaminated by various chemicals used from planting to processing. These chemicals have been shown to cause illnesses for which some concerned government agencies have instituted regulatory mechanisms to minimize the risks and the effects on humans. It is for these concerns that reliable and accurate rapid determination techniques are needed to effect proper regulatory standards for the protection of people's nutritional health. This paper, therefore, reports the comparative evaluation of the extraction methods in the determination of atrazine (commonly used in agricultural as a herbicide) residues in foods using supercritical fluid chromatography (SFC) and enzyme-linked immunosorbent assay (ELISA) techniques. Supercritical fluid extraction (SFE) and microwave solvent extraction (MSE) methods were used to test samples of frozen vegetables, fruit juice, and jam from local food markets in Houston. Results showed a high recovery percentage of atrazine residues using supercritical fluid coupled with ELISA and SFC than with MSE. Comparatively, however, atrazine was detected 90.9 and 54.5% using SFC and ELISA techniques, respectively. ELISA technique was, however, less time consuming, lower in cost, and more sensitive with low detection limit of atrazine residues than SFC technique. Mohamed H. El-Saeid, Ijeoma Kanu, Ebere C. Anyanwu, and Mahmoud A. Saleh Copyright © 2005 Mohamed H. El-Saeid et al. All rights reserved. Evaluation of DNA Damage in Common Carp (Cyprinus carpio L.) by Comet Assay for Determination of Possible Pollution in Lake Mogan (Ankara) Mon, 01 Jan 1900 00:00:00 +0000 Contamination of the aquatic environment with various concentrations of pollutants results in unexpected threats to humans and wildlife. The consequences of exposure and metabolism of pollutants/xenobiotics, especially carcinogens and mutagens, can be suitably assessed by investigating severe events, such as DNA damage; for example, DNA adducts and DNA strand breaks. One of the commonly used techniques to detect DNA damage in aquatic organisms is single-cell gel electrophoresis (comet assay). This study was carried out using Cyprinus carpio in order to identify the possible pollution in Lake Mogan, near Ankara, Turkey, where the city's sewer system and pesticides used in agriculture are believed to be the common causes of pollution. From the comet assay, the tail length (μm), tail intensity (%), and tail moment values of fish caught from Lake Mogan were found to be 31.10 ± 10.39, 7.77 ± 4.51, 1.50 ± 1.48, respectively, whereas for clean reference sites they were found to be 22.80 ± 1.08, 3.47 ± 1.59, 0.40 ± 0.51, respectively. The values are statistically different from each other (p < 0.0001, p < 0.0001, and p < 0.0013, respectively). These results indicate that Lake Mogan may be polluted with substances that have genotoxic effects and constitute an early warning for the lake system. Further detailed research is needed to establish the source of the pollution and the chemicals responsible. İsmet Çok, Onur Kenan Ulutaş, Öncü Okuşluk, Emre Durmaz, and Nilsun Demir Copyright © 2011 İsmet Çok et al. All rights reserved. Genotoxicity Studies in Groundwater, Surface Waters, and Contaminated Soil Mon, 01 Jan 1900 00:00:00 +0000 It is at present considered important to include biological tests in measuring programmes of environmental samples to supplement the chemical and physical parameters that are currently used. A battery of tests is therefore necessary, also within a given “endpoint” (e.g., genotoxicity), because one single test will not give all the answers to our questions. As it is not possible to include all available tests in routine screening programmes, a selection of tests should be made. According to comparative investigations, the bacterial Ames test remains very important. When no preconcentration step is involved, other bacterial tests (e.g., the umu-C and VITOTOXâ tests) may be recommended. The comet assay may be used in Daphnia or human white blood cells. Further validations, comparisons, mechanistic investigations, etc. remain necessary as differences are often found between the tests that are not solely explained by differences in genetic endpoint and that therefore should further be investigated and understood. Luc Verschaeve Copyright © 2002 Luc Verschaeve. All rights reserved. A dietary risk assessment of the pyrethroid insecticide resmethrin associated with its use for West Nile Virus mosquito vector control in California Mon, 01 Jan 1900 00:00:00 +0000 An outbreak of human illnesses associated with West Nile Virus (WNV) occurred in New York City in 1999. Since then, it has gradually spread westwards, reaching northern California for the first time in 2005. WNV is transmitted by several mosquito species and birds serve as the main reservoir. Several control measures have been used, targeting both the aquatic larvae and the adult mosquitoes. In the latter case, roosting birds in trees are sprayed with pyrethroid insecticides because these are highly toxic to mosquitoes, but have low avian toxicity. A request was made to use a resmethrin-containing insecticide during the month of October 2005 in California. Because resmethrin was not registered for use on growing crops, concerns were raised about potential crop contamination. Therefore, an expedited dietary risk assessment was conducted on resmethrin. Developmental toxicity in the rat (NOELs of 25 or 40 mg/kg/day) was used as the acute endpoint and dietary exposure was assessed using the DEEM-FCIDTM computer program. Only crops growing above ground during October were considered. Margins of Safety (MOS) were found to be above 100, the level generally considered to be sufficient to protect public health when using an animal NOEL. Wesley C. Carr, Poorni Iyer, and Derek W. Gammon Copyright © 2006 Jr., Wesley C. Carr et al. All rights reserved. A Bacterial Toxin that Causes DNA Damage to Modulate Cellular Responses Mon, 01 Jan 1900 00:00:00 +0000 Campylobacter jejuni constitutes the leading cause of bacterial diarrhea in the U.S. and all around the world [1]. This common bacterium produces a toxin known as cytolethal distending toxin (CDT) [2] which causes intoxicated cells to enlarge and to stop dividing with a double DNA content characteristic of G2/M arrest [3]. The effect of the toxin on the cell is so striking that it captivated scientists for a long time. However, its mechanism of action had remained elusive. Maria Lara-Tejero Copyright © 2001 Maria Lara-Tejero. All rights reserved. Human Exposure Assessment in Air Pollution Systems Mon, 01 Jan 1900 00:00:00 +0000 The air pollution problem can be depicted as a system consisting of several basic components: source, concentration, exposure, dose, and adverse effects. Exposure, the contact between an agent (e.g., an air pollutant) and a target (e.g., a human respiratory tract), is the key to linking the pollution source and health effects. Human exposure to air pollutants depends on exposure concentration and exposure duration. Exposure concentration is the concentration of a pollutant at a contact boundary, which usually refers to the human breathing zone. However, ambient concentrations of regulated pollutants at monitoring sites have been measured in practice to represent actual exposure. This can be a valid practice if the pollutants are ones that are predominantly generated outdoors and if the monitoring sites are appropriately selected to reflect where people are. Results from many exposure studies indicate that people are very likely to receive the greatest exposure to many toxic air pollutants not outside but inside places such as homes, offices, and automobiles. For many of these pollutants, major sources of exposure can be quite different from major sources of emission. This is because a large emission source can have a very small value of exposure effectiveness, i.e., the fraction of pollutant released from a source that actually reaches the human breathing zone. Exposure data are crucial to risk management decisions for setting priorities, selecting cost-effective approaches to preventing or reducing risks, and evaluating risk mitigation efforts. Measurement or estimate of exposure is essential but often inadequately addressed in environmental epidemiologic studies. Exposure can be quantified using direct or indirect measurement methods, depending upon the purpose of exposure assessment and the availability of relevant data. The rapidly developing battery and electronic technologies as well as advancements in molecular biology are expected to accelerate the improvement of current methods and the development of new methods for future exposure assessment. Junfeng (Jim) Zhang and Paul J. Lioy Copyright © 2002 Junfeng (Jim) Zhang and Paul J. Lioy. All rights reserved. In Vitro Exposure of Porcine Ovarian Follicular Cells to PCB 153 Alters Steroid Secretion But Not Their Viability — Preliminary Study Mon, 01 Jan 1900 00:00:00 +0000 In our previous paper[1], we demonstrated that porcine follicles collected during the early stage of development are the most sensitive to the toxic action of polychlorinated biphenyl 153 (PCB 153). Follicles of this type were collected to test the effect of PCB 153 on cell steroidogenesis and viability. Cocultures of granulosa and theca cells were grown in M199 medium at 37C. Control cultures were maintained in that medium alone, while experimental ones were supplemented with PCB 153 at doses of 5, 10, 50, and 100 ng/ml. After 48, 96, and 144 h, media were collected for steroid analysis and cell viability was measured using an LDH (lactate dehydrogenase activity) cytotoxicity test. A 2-day exposure of follicular cells to all the investigated doses of PCB 153 caused a statistically significant decrease in progesterone (P4) secretion, while in doses of 50 and 100 ng/ml there was also a decrease in testosterone (T) secretion. No effect on estradiol (E2) secretion was observed. The observed decrease in P4 and T secretion, and lack of any statistically significant effect on E2 secretion by cells from small follicles exposed for 48 h to PCB, suggests that PCB 153 acts before P4 formation. Longer exposures caused an increase in P4 secretion, with a concomitant drastic decrease in T secretion and a tendency to decrease the E2 secretion, suggesting inhibition of P450 17a hydroxysteroid dehydrogenase, an enzyme that converts P4 to T. The observed PCB 153induced increase in P4 secretion by cells collected from small antral follicles, with a concomitant decrease in E2 secretion, accounts for the induction of luteinization and, in this case, inhibition of aromatization process in the follicles. However, in all doses tested and at all times of exposure, PCB 153 had no effect on cell viability. These findings suggest different time of exposuredependent action of PCB 153 on particular steps of steroidogenesis but not action on cell viability. These results should be considered preliminary, pending confirmation by other studies. Ewa L. Gregoraszczuk and Anna K. Wjtowicz Copyright © 2002 Ewa L. Gregoraszczuk and Anna K. Wjtowicz. All rights reserved.